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Abstract:

Compounds according to formula (I) are effective for the treatment of
broncho-obstructive and inflammatory diseases.

Claims:

1. A compound of formula (I): ##STR00128## wherein: X is O or S; X' is
O or S; Y is NH or absent; R1 is aryl, heteroaryl,
aryl(C1-C6) alkyl, heteroaryl(C1-C6)alkyl, or a group
of formula (a) or (b): ##STR00129## wherein R3 and R4 are the
same or different and may be independently H or are selected from the
group consisting of (C3-C8)cycloalkyl, aryl,
aryl(C1-C6) alkyl, heteroaryl, and
heteroaryl(C1-C6)alkyl, each which may be optionally
substituted by one or more substituents selected from the group
consisting of a halogen atom, --OH, (C1-C6)haloalkyl,
(C1-C6)alkoxy, oxo (═O), --SH, --NO2, --CN,
--CONH2, --COOH, (C1-C6)alkoxycarbonyl,
(C1-C6)alkylsulfanyl, (C1-C6)alkylsulfinyl,
(C1-C6)alkylsulfonyl and (C1-C6)alkyl or, when
R3 and R4 are both independently aryl or heteroaryl they may be
linked to each other through a (CH2), group wherein r=0, 1, or 2, to
form a tricyclic ring system wherein any of the methylene
(CH2)r groups may be optionally replaced by a heteroatom or
heteroaromatic group selected from O, S, N and NH, and with the proviso
that R3 and R4 are not simultaneously H; R2 is a group of
formula (c) or (d): ##STR00130## wherein m=1, 2 or 3; n=1, 2 or 3;
A.sup.- is a physiologically acceptable anion; R5 is a group of
formula (e): --(CH2)p--P--(CH2)q--W (e) wherein p
is 0 or an integer from 1 to 4; q is 0 or an integer from 1 to 4; P is
absent or is selected from the group consisting of O, S, S(O)--,
--S(O2)--, --C(O)--, --CO(O)--, --N(R6)--, --CH═CH--,
--N(R6)(SO2)--, --N(R6)CO(O)--, --N(R6)C(O)--,
--SO2N(R6)--, --CO(O)N(R6)-- and --C(O)N(R6)--; W is
H or is selected from the group consisting of (C1-C6)alkyl,
(C2-C6)alkenyl, (C3-C8)cycloalkyl,
(C3-C8)heterocycloalkyl, aryl and heteroaryl, each of which is
optionally substituted by one or more substituents selected from the
group consisting of a halogen atom, --OH, oxo (═O), --SH, --NO2,
--N(R6)2, --CN, --CON(R6)2, --COOH, --NHCOR6,
CO2R6, (C1-C6)alkoxycarbonyl,
(C1-C6)alkylsulfanyl, (C1-C6)alkylsulfinyl,
(C1-C6)alkylsulfonyl, (C1-C6)alkyl,
(C1-C6)alkoxy, aryl, and heteroaryl; R6 is, independently
in each occurrence, H or is selected from the group consisting of
(C1-C6)alkyl, (C1-C6)haloalkyl,
(C2-C6)alkynyl, (C2-C6)alkenyl,
(C3-C8)cycloalkyl, heteroaryl, and aryl, each of which is
optionally substituted by one or more substituents selected from the
group consisting of halogen atoms, --OH, oxo (═O), --SH, --NO2,
--CN, --CONH2, --COOH, (C1-C6)alkoxycarbonyl,
(C1-C6)alkylsulfanyl, (C1-C6)alkylsulfinyl,
(C1-C6)alkylsulfonyl, (C3-C8)cycloalkyl,
(C1-C6)alkyl, and (C1-C6)alkoxy; or a
pharmaceutically acceptable salt thereof.

2. A compound or salt according to claim 1, which is a compound
represented by formula (IA): ##STR00131## wherein: R1 is a group
of formula (a), wherein R3 and R4 are the same or different and
are independently H or selected from the group consisting of aryl and
heteroaryl, each of which may be optionally substituted by one or more
substituents selected from the group consisting of a halogen atom,
(C1-C6)alkoxy, and (C1-C6)haloalkyl, and R2 is a
group of formula (c) or (d): ##STR00132## wherein n=m=1 and R5 is
a group of formula (e) wherein p is 0, 1, 2 or 3, P is absent or is
selected from the group consisting of --O--, --C(═O)--, and
--C(═O)N(H)--, q is 0, 1, or 2, and W is selected from the group
consisting of (C1-C6)alkyl, (C2-C6)alkenyl, aryl, and
heteroaryl, each of which is optionally substituted by one or more
substituents selected from the group consisting of a halogen atom, --OH,
--CN, (C1-C6)alkyl, and (C1-C6)alkoxy, or a
pharmaceutically acceptable salt thereof.

4. A compound or salt according to claim 1, which is a compound
represented by formula (IB): ##STR00133## wherein R1 is aryl or a
group of formula (a) or (b) wherein R3 and R4 are the same or
different and are independently H or selected from the group consisting
of aryl, aryl(C1-C6)alkyl, and heteroaryl, each which may be
optionally substituted by one or more substituents selected from the
group consisting of a halogen atom, (C1-C6)alkoxy, and
(C1-C6)haloalkyl or when R3 and R4 are both
independently aryl or heteroaryl they may be linked to each other through
a (CH2), group wherein r=0, 1, or 2, to form a tricyclic ring system
wherein any of the methylene (CH2)r groups may be substituted
by a heteroatom or heteroaromatic group selected from O, S, N and NH and
with the proviso that R3 and R4 are not simultaneously H, and
R2 is a group of formula (c) or (d): ##STR00134## wherein n=1 or
2, m=1 and R5 is a group of formula (e), wherein p=1 or 3, P is
absent or is selected from the group consisting of --O--, --C(═O)--
and --C(═O)O--, q=1 and W is selected from the group consisting of
(C1-C6)alkyl, (C2-C6)alkenyl, aryl, and heteroaryl
each of which is optionally substituted by one or more substituents
selected from the group consisting of halogen atoms, --OH, --CN,
(C1-C6)alkyl, (C1-C6)alkoxycarbonyl, and
(C1-C6)alkoxy.

6. A compound of formula (IV): ##STR00135## wherein Q represents a
group of formula (f) or (g): ##STR00136## wherein R7 is
(C1-C6)alkyl or aryl(C1-C6)alkyl, and R1, X, X',
n, m and Y are as defined in claim 1, or a pharmaceutically acceptable
salt thereof.

7. A compound or salt according to claim 6, which is a compound
represented by formula (IVA): ##STR00137## wherein R1 is a group
of formula (a): ##STR00138## R3 and R4 are the same or
different and are independently H or selected from the group consisting
of aryl and heteroaryl, each of which may be optionally substituted by
one or more substituents selected from the group consisting of a halogen
atom, (C1-C6)alkoxy, and (C1-C6)haloalkyl; and Q is a
group of formula (f) or (g): ##STR00139## wherein R7 is
aryl(C1-C6)alkyl, or a pharmaceutically acceptable salt
thereof.

8. A compound or salt according to claim 6, which is a compound
represented by formula (IVB): ##STR00140## wherein R1 is aryl or a
group of formula (a), wherein R3 and R4 are the same or
different and are independently H or selected from the group consisting
of aryl, aryl(C1-C6)alkyl, and heteroaryl, each of which may be
optionally substituted by one or more substituents selected from the
group consisting of halogen atoms, (C1-C6)alkoxy, and
(C1-C6)haloalkyl or when R3 and R4 are both
independently aryl or heteroaryl they may be linked to each other through
a (CH2), group wherein r=0, 1, or 2, to form a tricyclic ring system
wherein any of the methylene (CH2), groups may be a heteroatom or
heteroaromatic group selected from O, S, N and NH, and with the proviso
that R3 and R4 are not simultaneously H; and Q is a group of
formula (f) or (g): ##STR00141## wherein n is l, m is 2 and R5 is
a group of formula (f) wherein p=0, P is absent, q is 0 or 1 and W is
(C1-C6)alkyl, or a pharmaceutically acceptable salt thereof.

9. A pharmaceutical composition, comprising a compound or salt according
to claim 1 and one or more pharmaceutically acceptable carriers and/or
excipients.

10. A combination of a compound or salt according to claim 1 and one or
more active ingredients selected from the group consisting of a
beta2-agonist, a corticosteroid, a P38 MAP kinase inhibitor, a IKK2
inhibitor, a HNE inhibitor, a PDE4 inhibitor, aleukotriene modulator, a
NSAID, and a mucus regulator.

11. A pharmaceutical composition according to claim 9, which is in a form
suitable for administered by inhalation.

12. A pharmaceutical composition according to claim 11, which is an
inhalable powder, a propellant-containing metering aerosol, or a
propellant-free inhalable formulation.

13. A device, comprising a pharmaceutical composition according to claim
11, wherein said device is a single- or multi-dose dry powder inhaler, a
metered dose inhaler, or a soft mist nebulizer.

14. A method for treating a broncho-obstructive or inflammatory disease,
comprising administering an effective amount of a compound or salt
according to claim 1 to a subject in need thereof.

15. A method according to claim 14, wherein said disease is asthma,
chronic bronchitis, or chronic obstructive pulmonary disease.

Description:

CROSS REFERENCES TO RELATED APPLICATIONS

[0001] This application claims priority to European Patent Application No.
11164337.5, filed on Apr. 29, 2011, which is incorporated herein by
reference in its entirety.

BACKGROUND OF THE INVENTION

[0002] 1. Field of the Invention

[0003] The present invention relates to compounds which act as muscarinic
receptor antagonists, to methods of preparing such compounds, to
compositions which contain such a compound, and therapeutic uses of such
compounds.

[0004] 2. Discussion of the Background

[0005] Quaternary ammonium salts acting as muscarinic (M) receptor
antagonist drugs are currently used in therapy to induce bronchodilation
for the treatment of respiratory diseases. Examples of well known M
receptor antagonists are for instance represented by ipratropium bromide
and tiotropium bromide.

[0006] Some chemical classes acting as selective M3 receptor antagonist
drugs have been developed for the treatment of inflammatory or
obstructive airway diseases such as asthma and chronic obstructive
pulmonary disease (COPD).

[0007] Quinuclidine carbamate derivatives and their use as M3 antagonists
are for instance disclosed in WO 02/051841, WO 03/053966, and WO
2008/012290, which are incorporated herein by reference in their
entireties. WO 2010/015324, which is incorporated herein by reference in
its entirety, describes carbonate derivatives and their use as M3
antagonists.

[0008] There remains, however, a need for compounds characterized by a
good activity as M3 antagonists and by an improved pulmonary stability.

SUMMARY OF THE INVENTION

[0009] Accordingly, it is one object of the present invention to provide
novel compounds which act as muscarinic receptor antagonists.

[0010] It is another object of the present invention to provide novel
methods of preparing such a compound.

[0011] It is another object of the present invention to provide novel
compositions which contain such a compound.

[0012] It is another object of the present invention to provide novel
therapeutic uses of such a compounds.

[0013] These and other objects, which will become apparent during the
following detailed description, have been achieved by the inventors'
discovery that compounds of general formula (I), described below, are
characterized by a good activity as M3 antagonists and by an improved
pulmonary stability.

[0015] The present invention also provides processes for the preparation
of such a compound.

[0016] The present invention also provides compositions which contain such
a compound.

[0017] The present invention also provides therapeutic uses of such a
compound.

[0018] The present invention also provides combinations of such a compound
with other pharmaceutical active ingredients, for instance, those
currently used in the treatment of respiratory disorders, e.g.
beta2-agonists, corticosteroids, P38 MAP kinase inhibitors, IKK2, HNE
inhibitors, PDE4 inhibitor, leukotriene modulators, NSAIDs, and mucus
regulators.

[0019] The compounds of the present invention are characterized by a good
activity as M3 antagonists and by an improved pulmonary stability.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0020] Thus, in a first embodiment, the present invention provides novel
compounds of general formula (I):

##STR00001##

wherein:

[0021] X may be O or S;

[0022] X' may be O or S;

[0023] Y may be NH or absent;

[0024] R1 is aryl, heteroaryl, aryl(C1-C6)alkyl,
heteroaryl(C1-C6)alkyl, or a group of formula (a) or (b):

##STR00002##

[0025] wherein [0026] R3 and R4 are the same or different and
may be independently H or are selected from the group consisting of
(C3-C8)cycloalkyl, aryl, aryl(C1-C6)alkyl,
heteroaryl, and heteroaryl(C1-C6)alkyl, which may be optionally
substituted by one or more substituents selected from the group
consisting of halogen atoms, --OH, (C1-C6)haloalkyl,
(C1-C6)alkoxy, oxo (═O), --SH, --NO2, --CN,
--CONH2, --COOH, (C1-C6)alkoxycarbonyl,
(C1-C6)alkylsulfanyl, (C1-C6)alkylsulfinyl,
(C1-C6)alkylsulfonyl, and (C1-C6)alkyl or, when
R3 and R4 are both independently aryl or heteroaryl they may be
linked to each other through a (CH2), with r=0-2, to form a
tricyclic ring system wherein any of the methylene (CH2), groups may
be optionally replaced by a heteroatom or heteroaromatic group selected
from O, S, N, and NH, and with the proviso that R3 and R4 are
not simultaneously H;

[0032] (CH2)p--P--(CH2)q--W (e) [0033] wherein
[0034] p is 0 or an integer from 1 to 4; [0035] q is 0 or an integer from
1 to 4; [0036] P is absent or is selected from the group consisting of
--O--, --S--, --S(O)--, --S(O2)--, --C(O)--, --CO(O)--,
--N(R6)--, --CH═CH--, --N(R6)(SO2)--,
--N(R6)CO(O)--, --N(R6)C(O)--, --SO2N(R6)--,
--CO(O)N(R6)-- and --C(O)N(R6)--; [0037] W is selected from the
group consisting of H, (C1-C6)alkyl, (C2-C6)alkenyl,
(C3-C8)cycloalkyl, (C3-C8)heterocycloalkyl, aryl, and
heteroaryl, optionally substituted by one or more substituents selected
from the group consisting of halogen atoms, --OH, oxo (═O), --SH,
--NO2, --N(R6)2, --CN, --CON(R6)2, --COOH,
--NHCOR6, --CO2R6, (C1-C6)alkoxycarbonyl,
(C1-C6)alkylsulfanyl, (C1-C6)alkylsulfinyl,
(C1-C6)alkylsulfonyl, (C1-C6)alkyl,
(C1-C6)alkoxy, aryl, and heteroaryl; [0038] R6 is,
independently in each occurrence, H or is selected from the group
consisting of (C1-C6)alkyl, (C1-C6)haloalkyl,
(C2-C6)alkynyl, (C2-C6)alkenyl,
(C3-C8)cycloalkyl, heteroaryl, and aryl, optionally substituted
by one or more substituents selected from the group consisting of halogen
atoms, --OH, oxo (═O), --SH, --NO2, --CN, --CONH2, --COOH,
(C1-C6)alkoxycarbonyl, (C1-C6)alkylsulfanyl,
(C1-C6)alkylsulfinyl, (C1-C6)alkylsulfonyl,
(C3-C8)cycloalkyl, (C1-C6)alkyl and
(C1-C6)alkoxy; and pharmaceutically acceptable salts thereof.

[0039] The present invention is also directed to compounds of general
formula (IV):

##STR00004##

wherein Q represents a group of formula (f) or (g):

##STR00005##

R7 is selected from the group consisting of (C1-C6)alkyl
and aryl(C1-C6)alkyl, and R1, X, X', n, m and Y have the
above reported meanings in formula (I).

[0040] The term "halogen atoms" as used herein includes fluorine,
chlorine, bromine, and iodine.

[0041] The expression "(C1-C6)alkyl" refers to straight-chained
or branched alkyl groups wherein the number of carbon atoms is from 1 to
6. Examples of groups are methyl, ethyl, n-propyl, isopropyl, t-butyl,
pentyl, hexyl and the like.

[0044] The expressions "(C1-C6)haloalkyl" and
"(C1-C6)haloalkoxy" refer to the above "(C1-C6)alkyl"
and "(C1-C6)alkoxy" groups wherein one or more hydrogen atoms
are replaced by one or more halogen atoms, which can be the same or
different from each other.

[0046] The expression "(C2-C6)alkenyl" refers to straight or
branched carbon chains with one or more double bonds. Examples of said
groups may thus comprise ethenyl, propenyl, butenyl, pentenyl, hexenyl
and the like.

[0047] The expression "(C2-C6)alkynyl" refers to straight or
branched carbon chains with one or more triple bonds. Examples of said
groups may thus comprise ethinyl, propinyl, butinyl, pentinyl, hexinyl,
and the like.

[0048] The expression "(C3-C8)cycloalkyl" refers to mono or
bi-cycloaliphatic hydrocarbon groups with from 3 to 8 carbon atoms.
Examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,
cycloheptyl, bicyclo[2.2.1]hept-2-yl, and the like.

[0049] The derived expression "(C3-C8)heterocycloalkyl" refers
to (C3-C8)cycloalkyl groups, in which at least one ring carbon
atom is replaced by a heteroatom or heteroaromatic group (e.g. N, NH, S,
or O). Examples include quinuclidinyl, pyrrolidinyl, piperidinyl and the
like.

[0050] The expression "aryl" refers to mono-, bi-, or tricyclic ring
systems having 5 to 20, preferably from 5 to 15, ring atoms, and wherein
at least one ring is aromatic.

[0051] The expression "heteroaryl" refers to mono-, bi- or tri-cyclic ring
systems with 5 to 20 ring atoms, preferably from 5 to 15, in which at
least one ring is aromatic and in which at least one carbon ring atom is
a heteroatom or heteroaromatic group (e.g. N, NH, S or O).

[0057] Besides the presence of A.sup.- anion, whenever further basic amino
groups are present in the compounds of formula (I), additional
physiological acceptable anions, among those formerly indicated, may be
present. Likewise, in the presence of acidic groups such as --COOH
groups, corresponding physiological cation salts may be present as well,
for instance including alkaline or alkaline earth metal ions.

[0058] In the present description, and unless otherwise provided, within
formula (I) or (IV), Y may represent a divalent --NH-- group or,
whenever, absent, it clearly represents a single bond so as to provide
compounds R1--X--C(═X')R2 or R1--X--C(═X')Q.

[0059] Furthermore, when both R3 and R4 are independently
selected from aryl or heteroaryl groups, the said groups may be linked to
each other through a (CH2), group, so as to form a tricyclic ring
system.

[0060] From all the above, it is clear to the skilled person that, when r
is 0, the said R3 and R4 groups are linked to each other
through a bond.

[0061] Unless otherwise provided, within formula (I) and (IV), R2 and
Q groups being represented by groups (c), (d) and (f), (g), the asterisks
in these latter represent their point of attachment to the rest of the
molecule.

[0062] As an example, the following groups can be highlighted:

##STR00006##

[0063] It will be apparent that the compounds of general formula (I) and
(IV) may contain asymmetric centers. Therefore the invention also
includes the optical stereoisomers and mixtures thereof.

[0064] Where the compounds according to the present invention possess two
or more asymmetric centers, they may additionally exist as
diastereoisomers. It is to be understood that all such isomers and
mixtures thereof in any proportion are encompassed within the scope of
the present invention.

[0065] More in particular, the active compounds of formula (I) and (IV)
show at least one chiral center, which is represented by the carbon atom
in Q or R2 and which is directly linked to group Y.

[0066] Therefore, according to a specific embodiment, in compound (I), the
carbon atom of R2 group being linked to Y is in the form of
(S)-enantiomer when R2 is a group of formula (c).

[0067] According to a preferred embodiment, in compound (I), the carbon
atom of R2 group being linked to Y is in the form of (R)-enantiomer
when R2 is a group of formula (c).

[0068] According to another embodiment, in compound (IV), the carbon atom
of Q group being linked to Y is in the form of (S)-enantiomer when Q is a
group of formula (f), or a group of formula (g), in this latter case
obviously except when m is 2 and n is l, m is 3 and n is 2.

[0069] According to a preferred embodiment, in compound (IV), the carbon
atom of Q group being linked to Y is in the form of (R)-enantiomer when Q
is a group of formula (f), or a group of formula (g), in this latter case
obviously except when m is 2 and n is l, m is 3 and n is 2.

[0070] In the compounds of general formula (I) of the present invention,
when R2 is a group of formula (a) and R3 and R4 have
different meanings, the carbon atom bearing R3 and R4 is a
chiral center.

[0071] Moreover, within the compounds of formula (I), when R1
represents a group of formula (b), R3 is bound to the carbon atom
bearing a double bond in any of the possible (Z) or (E) configurations
and presently identified through the symbol

##STR00007##

[0072] A first preferred group of compounds is that of general formula
(IVA):

##STR00008##

wherein R1 is a group of formula (a):

##STR00009##

wherein R3 and R4 are the same or different and are H or
selected from the group consisting of aryl and heteroaryl, which may be
optionally substituted by one or more substituents selected from the
group consisting of halogen atoms, (C1-C6)alkoxy, and
(C1-C6)haloalkyl, Q is a group of formula (f) or (g):

##STR00010##

wherein n=m=1 and R5 is a group of formula (e):

--(CH2)p-P--(CH2)q-W (e)

wherein p=0, P is absent, q=1 and W is aryl.

[0073] Still more preferred, within this class, are the compounds of
general formula (IVA) wherein R1 is selected from
bis(3-fluorophenyl)methyl, benzhydryl, (4-methoxyphenyl)(phenyl)methyl,
(2-fluorophenyl)(4-fluorophenyl)methyl,
(2-fluorophenyl)(3-fluorophenyl)methyl,
((3,4-difluorophenyl)(phenyl)methyl, 4-(trifluoromethyl)phenyl)methyl,
(2-chlorophenyl)(4-chlorophenyl)methyl, and thiophen-2-ylmethyl, and Q is
selected from quinuclidinyl and benzylpyrrolidinyl.

[0074] A second preferred group of compounds is that of general formula
(IA):

##STR00011##

wherein R1 is a group of formula (a):

##STR00012##

wherein R3 and R4 are the same or different and are H or
selected from the group consisting of aryl and heteroaryl, which may be
optionally substituted by one or more substituents selected from the
group consisting of halogen atoms, (C1-C6)alkoxy, and
(C1-C6)haloalkyl, R2 is a group of formula (c) or (d):

##STR00013##

wherein n=m=1 and R5 is a group of formula (e) wherein p is 0, 1, 2
or 3, P is absent or is selected from the group consisting of --O--,
--C(═O)-- and --CONH--, q is 0, 1 or 2, and W is selected from the
group consisting of (C1-C6)alkyl, (C2-C6)alkenyl,
aryl, and heteroaryl optionally substituted by one or more substituents
selected from the group consisting of halogen atoms, --OH, --CN,
(C1-C6)alkyl, and (C1-C6)alkoxy.

[0076] A third preferred group of compounds is that of general formula
(IVB):

##STR00014##

wherein R1 is aryl or a group of formula (a):

##STR00015##

R3 and R4 are the same or different and are H or are selected
from the group consisting of aryl, aryl(C1-C6)alkyl, and
heteroaryl, which may be optionally substituted by one or more
substituents selected from the group consisting of halogen atoms,
(C1-C6)alkoxy, and (C1-C6)haloalkyl or when R3
and R4 are both independently aryl or heteroaryl they may be linked
to each other through a (CH2)r with r=0-2, to form a tricyclic
ring system wherein any of the methylene (CH2)r groups may be a
heteroatom or heteroaromatic group selected from O, S, N, and NH, and
with the proviso that R3 and R4 are not simultaneously H; Q is
a group of formula (f) or (g):

##STR00016##

wherein n is l, m is 2 and R7 is a group of formula (f) wherein p=0,
P is absent, q is 0 or 1 and W is (C1-C6)alkyl.

[0077] Still more preferred, within this class, are the compounds of
general formula (IVB) wherein R1 is selected from
(3,4-difluorophenyl)(phenyl)methyl, bis(3-fluorophenyl)methyl,
1,2-diphenylethyl, bis(4-chlorophenyl)methyl, bis(4-fluorophenyl)methyl,
benzhydryl, (4-methoxyphenyl)(phenyl)methyl,
(2-chlorophenyl)(4-chlorophenyl)methyl, 1,2-diphenylvinyl,
3-fluorobenzyl, benzyl, and fluorenyl, and Q is selected from
quinuclidin-3-yl, and methylpiperidinyl.

[0078] A fourth preferred group of compounds is that of general formula
(IB):

##STR00017##

wherein R1 is aryl or a group of formula (a) or (b):

##STR00018##

wherein R3 and R4 are the same or different and are H or
selected from the group consisting of aryl, aryl(C1-C6)alkyl,
and heteroaryl, which may be optionally substituted by one or more
substituents selected from the group consisting of halogen atoms,
(C1-C6)alkoxy, and (C1-C6)haloalkyl or when R3
and R4 are both independently aryl or heteroaryl they may be linked
to each other through a (CH2)r with r=0-2, wherein when n=0, to
form a tricyclic ring system wherein any of the methylene
(CH2)r may be a heteroatom or heteroaromatic group selected
from O, S, N and NH, and with the proviso that R3 and R4 are
not simultaneously H, R2 is a group of formula (c) or (d):

##STR00019##

wherein n=1 or 2, m=1 and R5 is a group of formula (e), wherein p=1
or 3, P is absent or is selected from the group consisting of --O--,
--C(═O)-- and --C(═O)O--, q=1 and W is selected from the group
consisting of (C1-C6)alkyl, (C2-C6)alkenyl, aryl, and
heteroaryl optionally substituted by one or more substituents selected
from the group consisting of halogen atoms, OH, CN,
(C1-C6)alkyl, (C1-C6)alkoxycarbonyl, and
(C1-C6)alkoxy.

[0080] A fifth preferred group of compounds is that of general formula
(IC):

##STR00020##

wherein R1 is a group of formula (a) wherein R3 and R4 are
independently aryl groups, which may be optionally substituted by one or
more halogen atoms, R2 is a group of formula (c):

##STR00021##

wherein R5 is a group of formula (e) with p=1, P is --C(═O)--, q
is 0, and W is heteroaryl.

[0081] Still more preferred, within this class, are the compounds of
general formula (IC) wherein R1 is bis(3-fluorophenyl)methyl and
R2 is (2-oxo-2-(thiophen-2-yl)ethyl)-1-azoniabicyclo[2.2.2]octanyl.

[0082] A sixth preferred group of compounds is that of general formula
(IVC):

##STR00022##

wherein R1 is a group of formula (a) wherein R3 and R4 are
independently aryl groups, optionally substituted by one or more halogen
atoms, Q is a group of formula (f):

##STR00023##

[0083] Still more preferred, within this class, are the compounds of
general formula (IVC) wherein R1 is bis(3-fluorophenyl)methyl and Q
is quinuclidin-3-yl.

[0084] A seventh preferred group of compounds is that of general formula
(ID):

##STR00024##

wherein R1 is a group of formula (a) wherein R3 and R4 are
independently aryl groups, which may be optionally substituted by one or
more halogen atoms, R2 is a group of formula (c):

##STR00025##

wherein R5 is a group of formula (e) with p=1, P is --C(═O)--, q
is 0, and W is heteroaryl.

[0085] Still more preferred, within this class, are the compounds of
general formula (ID) wherein R1 is benzyl and R2 is
(2-oxo-2-(thiophen-2-yl)ethyl)-1-azoniabicyclo[2.2.2]octanyl.

[0086] An eighth preferred group of compounds is that of general formula
(IVD):

##STR00026##

wherein R1 is a group of formula (a) wherein R3 and R4 are
independently aryl groups, optionally substituted by one or more halogen
atoms, Q is a group of formula (f):

##STR00027##

[0087] Still more preferred, within this class, are the compounds of
general formula (IVC) wherein R1 is benzyl and Q is quinuclidinyl.

[0088] The present invention also provides pharmaceutical compositions of
compounds of formula (I) or (IV) alone or in combination with or in
admixture with one or more pharmaceutically acceptable carriers and/or
excipients.

[0089] The present invention also provides the use of compounds of formula
(I) or (IV) for preparing a medicament.

[0090] In a further aspect, the present invention provides the use of
compounds of formula (I) or (IV) for the prevention and/or treatment of
any broncho-obstructive or inflammatory disease, preferably asthma or
chronic bronchitis or chronic obstructive pulmonary disease (COPD).

[0091] In a further aspect, the present invention provides the use of
compounds of formula (I) or (IV) for the manufacture of a medicament for
the prevention and/or treatment of any broncho-obstructive or
inflammatory disease, preferably asthma or chronic bronchitis or chronic
obstructive pulmonary disease (COPD).

[0092] The present invention further provides a method for prevention
and/or treatment of any broncho-obstructive or inflammatory disease,
preferably asthma or chronic bronchitis or chronic obstructive pulmonary
disease (COPD), which comprises administering to a subject in need
thereof a therapeutically effective amount of a compound of general
formula (I) or (IV).

[0093] The present invention also provides pharmaceutical compositions
suitable for administration by inhalation.

[0095] The invention is also directed to a device which may be a single-
or multi-dose dry powder inhaler, a metered dose inhaler and a soft mist
nebulizer comprising the compounds of formula (I) or (IV).

[0096] The invention is also directed to a kit comprising the
pharmaceutical compositions of compounds of formula (I) or (IV) alone or
in combination with or in admixture with one or more pharmaceutically
acceptable carriers and/or excipients and a device which may be a single-
or multi-dose dry powder inhaler, a metered dose inhaler or a soft mist
nebulizer comprising the compounds of general formula (I) or (IV).

[0097] The present invention is also directed to a process for the
preparation of compounds of general formula (IV) and (I), which process
comprises:

[0098] (a) reacting a compound of general formula (II):

##STR00028##

wherein z is a carboxyl group or its corresponding acyl chloride
derivative, or an amino group and Q is as defined above, with a compound
of general formula (III):

##STR00029##

wherein L is H or an alkaline or alkaline earth metal, R1 and X are
as defined above, to obtain a compound of general formula (IV):

##STR00030##

wherein X', Q and Y are as described above, the reaction being carried
out in the presence of a suitable amount(s) of a condensing agent(s); and
optionally,

[0099] (b) alkylating the compound of general formula (IV) with an agent
of general formula (VI):

R5-A (VI)

wherein R5 and A are as described above, to give compounds of
general formula (I):

##STR00031##

[0100] According to specific embodiments, the present invention provides
the compounds reported below:

[0101] Compounds of general formula (I) and (IV) may be prepared according
to the following synthetic Scheme 1.

##STR00032##

[0102] Unless otherwise noted, R1, R2, Q, R5, X, X', Y, and
A are as defined above. It is well understood that protecting groups for
sensitive or reactive groups are employed where necessary in accordance
with general principles of chemistry. Protecting groups are manipulated
according to standard methods of organic synthesis (see Green T. W. and
Wuts P. G. M. (1991) Protecting Groups in Organic Synthesis, John Wiley
et Sons, which is cincroporated herein by reference in its entirety).

General Procedure for the Preparation of Compounds of Formula (I) and
(IV).

[0103] Compounds of general formula (IV), reported in the present
invention, may be prepared starting from compounds of general formula
(II), in which z may be either a carboxyl or an amino group and Q may be
a group of general formula (f) or (g):

##STR00033##

wherein m, n and R5 are as described above.

[0104] These compounds (II) can be reacted with compounds of general
formula (III), in which R1 and X are as described above and L can be
H or an alkaline or alkaline earth metal (i.e. potassium, lithium,
sodium, calcium and so on).

[0105] Starting materials of general formula (II) and (III) are
commercially available or may be conveniently prepared according to
standard procedures extensively reported in literature.

[0106] When z is a carboxyl group, the coupling between compounds of
general formula (II) and (III) may be promoted using standard amidation
and peptide coupling conditions. The operative conditions are chosen on
the basis of the reactivity of the acid (II) over alcohol/thioalcohol
(III) and of the compatibility of other groups being present in both
compounds (for a general reference on the above reaction and operative
conditions thereof see, for instance, Carey, F. A. and Sundeberg, R. J.
Advanced Organic Chemistry, Third Edition (1990), Plenum Press, New York
and London, pg 145), which is incorporated herein by reference in its
entirety.

[0107] The said conditions include, for instance, activating acid (II) by
means of one or more equivalents of a commercially available condensing
agent such as a carbodiimide (e.g. 1-ethyl-3-(3-dimethylaminopropyl)
carbodiimide (EDC) and the like) for example in the presence of
N-hydroxybenzotriazole (HOBt) followed by reaction of the activated
intermediate with alcohol or thioalcohol (III). An organic base such as
triethylamine and the like may be also present in the reaction mixture.
The activated intermediate may be either isolated, or pre-formed or
generated in situ, and then properly reacted with compounds of formula
(III). Suitable solvents for the coupling reaction include, but are not
limited to, halocarbon solvents (e.g. dichloromethane (DCM)),
tetrahydrofuran (THF), dioxane, dimethylformamide (DMF), and
acetonitrile. The reaction proceeds at temperature ranging from about
0° C. up to about 170° C., for a time period in the range
of about 1 hour up to about 96 hours. The reaction may be carried out
under conventional heating (using an oil bath) or under microwave
irradiation. The reaction may be conducted either in an open vessel or in
a sealed tube.

[0108] In some embodiments of the invention, acid (II) can be most
conveniently activated as acyl halide such as acyl chloride (z=COCl).
This activation may be affected according to one of the several standard
procedures reported in the literature. They comprise, for instance,
treatment of acid (II) with one or more equivalents of oxalyl chloride or
thionyl chloride. This reaction may be conducted in the presence of a
catalytic amount of dimethylformamide (DMF) in a suitable solvent (e.g.
dichloromethane) or neat, at temperature ranging from about 0° C.
to about 120° C. The activated intermediate may be either
isolated, or pre-formed or generated in situ. This intermediate can then
be reacted with alcohol or thioalcohol (III), using known methods in
order to obtain compounds of formula (IV). The reaction may be promoted
by a base such as triethylamine, pyridine and 4-dimethylaminopyridine,
and carried out in a suitable solvent (e.g. dichloromethane) or neat.
This reaction is performed in a temperature range from about 0° C.
to about 140° C. over a period of about 1 hour to about 74 hours.
The reaction may be conducted under conventional heating (using an oil
bath) or under microwave heating. The reaction may be carried out in an
open vessel or in a sealed tube.

[0109] The resulting esters (X, X'═O) and thioesters (X═S;
X'═O) of general formula (IV) can be then alkylated to obtain
compounds of general formula (I). Alternatively, they can be first
transformed into the corresponding thienoester (X═O; X'═S) or
di-thioester (X, X'═S) of general formula (IV). This conversion can
be effected according to one of the known standard procedures. For
instance, esters (IV) can be treated with Lawessons's reagent (see
Nicolaou, K. C. et al., Journal of the American Chemical Society, 1990,
12/17, pp. 6263-6276, which is incorporated herein by reference in its
entirety) or with tetraphosphorus decasulfide (see Cho, D. et al.,
Tetrahedron, 2010, 66/30, pp. 5583-5588, which is incorporated herein by
reference in its entirety) to achieve the corresponding thienoester (IV).
Likewise, thioesters may be converted into di-thioester by treatment with
Lawessons's reagent (see Cohen, O. et al., Tetrahedron, 2010, 66/20, pp.
3579-3582, which is incorporated herein by reference in its entirety).
The resulting thienoester (X═O; X'═S) or di-thioester (X,
X'═S) of general formula (IV) can then be alkylated to obtain
compounds of general formula (I).

[0110] When z is an amino group, compound of general formula (II) may be
coupled with compound of general formula (III) to afford carbamate,
thiocarbamate or dithiocarbamate of general formula (IV). This coupling
may be effected according to one of the standard procedures broadly
reported in the literature (a survey of the suitable reactions is given
by Chaturvedi, D. Current Organic Synthesis, 2007, 3, 308 or by Smith, M.
B. and March, J., March's Advanced Organic Chemistry, Fifth Edition
(2001), John Wiley & Sons, Inc., New York, appendix B, 1660, both of
which are incorporated herein by reference in their entireties). For
instance, the amine (II) could be treated with suitable activating
reagent that could be selected from, but are not limited to,
1,1'-carbonyldiimidazole, 1,1'-thiocarbonyldiimidazole, diphosgene,
triphosgene, or p-nitrophenylchloroformate. The reaction may be promoted
by a base selected from the group consisting of triethylamine, pyridine,
4-dimethylaminopyridine, and the like, in a suitable solvent (e.g.
dimethylformamide (DMF), tetrahydrofuran (THF), dichloromethane (DCM)).
The activated intermediate is generally pre-formed but it may be either
generated in situ or isolated. Then, the activated amine is reacted with
an alcohol or thioalcohol of formula (III), most conveniently dissolved
in the same solvent used for the activation of compound (II). Preferably,
the alcohol or thioalcohol is preliminary treated with a base
advantageously selected from NaH, BuLi (butyl lithium), and lithium
diisopropylamide (LDA).

[0111] The compounds of general formula (IV) wherein Q is a group of
formula (f) or (g), are eventually alkylated with an agent of general
formula (VI) to give compounds of general formula (I), wherein R2 is
a group with the above described meanings.

[0112] This kind of reaction is largely described in literature under
several different conditions. For instance, the reaction may be performed
neat or in a suitable solvent selected from the group consisting of
acetonitrile, ethyl acetate, DMF and tetrahydrofuran. The reaction
typically proceeds at temperature range from about 0° C. up to
about 170° C., for a time in the range of few minutes up to about
72 hours. The reaction may be carried out under conventional heating
(using an oil bath) or under microwave irradiation. The reaction may be
conducted either in an open vessel or in a sealed tube.

[0113] Compounds of general formula (I) can be either considered as final
products or can be further reacted to prepare other compounds of general
formula (I). Thus, any suitable moiety of R1 or R2 group in
general formula (I) could undergo a variety of reactions, to afford other
final compounds of general formula (I).

[0114] Likewise, the optional salification of the compounds of formula (I)
may be carried out by properly converting any of the free acidic groups
(e.g. carboxylic) or free amino groups into the corresponding
pharmaceutically acceptable salts.

[0115] In this case too, the operative conditions used for the optional
salification of the compounds of the invention are conventional

[0116] Further, depending from any of the meanings provided to R1 and
R2, among those reported above, it will be clear to the skilled
person that asymmetric centers may be present within the compounds of
formula (I). Therefore, the invention also includes any of the optical
stereoisomers, diastereoisomers, and mixtures thereof, in any proportion.

[0117] The present invention also provides pharmaceutical compositions of
compounds of formula (I) in admixture with one or more pharmaceutically
acceptable carriers, for example those described in Remington's
Pharmaceutical Sciences Handbook, XVII Ed., Mack Pub., N.Y., U.S.A.,
which is incorporated herein by reference in its entirety.

[0118] Administration of the compounds of the present invention may be
accomplished according to patient needs, for example, orally, nasally,
parenterally (subcutaneously, intravenously, intramuscularly,
intrasternally, and by infusion), by inhalation, rectally, vaginally,
topically, locally, transdermally, and by ocular administration.

[0119] Various solid oral dosage forms can be used for administering
compounds of the present invention including such solid forms as tablets,
gelcaps, capsules, caplets, granules, lozenges, and bulk powders. The
compounds of the present invention can be administered alone or combined
with various pharmaceutically acceptable carriers, diluents (such as
sucrose, mannitol, lactose, starches) and known excipients, including
suspending agents, solubilizers, buffering agents, binders,
disintegrants, preservatives, colorants, flavorants, lubricants, and the
like. Time release capsules, tablets and gels are also advantageous in
administering the compounds of the present invention.

[0120] Various liquid oral dosage forms can also be used for administering
compounds of the present invention, including aqueous and non-aqueous
solutions, emulsions, suspensions, syrups, and elixirs. Such dosage forms
can also contain suitable known inert diluents such as water and suitable
known excipients such as preservatives, wetting agents, sweeteners,
flavorants, as well as agents for emulsifying and/or suspending the
compounds of the invention. The compounds of the present invention may be
injected, for example, intravenously, in the form of an isotonic sterile
solution. Other preparations are also possible.

[0121] Suppositories for rectal administration of the compounds of the
present invention can be prepared by mixing the compound with a suitable
excipient such as cocoa butter, salicylates, and polyethylene glycols.

[0122] Formulations for vaginal administration can be in the form of
cream, gel, paste, foam, or spray formula containing, in addition to the
active ingredient, such as suitable carriers, are also known.

[0123] For topical administration, the pharmaceutical composition can be
in the form of creams, ointments, liniments, lotions, emulsions,
suspensions, gels, solutions, pastes, powders, sprays, and drops suitable
for administration to the skin, eye, ear, or nose. Topical administration
may also involve transdermal administration via means such as transdermal
patches.

[0124] For the treatment of the diseases of the respiratory tract, the
compounds according to the invention are preferably administered by
inhalation.

[0126] For administration as a dry powder, known single- or multi-dose
inhalers may be utilized. In that case the powder may be filled in
gelatine, plastic, or other capsules, cartridges or blister packs or in a
reservoir.

[0127] A diluent or carrier, generally non-toxic and chemically inert to
the compounds of present the invention, e.g. lactose or any other
additive suitable for improving the respirable fraction may be added to
the powdered compounds of the invention.

[0128] Inhalation aerosols containing propellant gas such as
hydrofluoroalkanes may contain the compounds of the present invention
either in solution or in dispersed form. The propellant-driven
formulations may also contain other ingredients such as co-solvents,
stabilizers, and optionally other excipients.

[0129] The propellant-free inhalable formulations comprising the compounds
of the present invention may be in form of solutions or suspensions in an
aqueous, alcoholic, or hydroalcoholic medium, and they may be delivered
by known jet or ultrasonic nebulizers or by soft-mist nebulizers such as
Respimat®.

[0130] The compounds of the present invention may be administered as the
sole active agent or in combination with one or more other pharmaceutical
active ingredients including those currently used in the treatment of
respiratory disorders, e.g. beta2-agonists, corticosteroids and
anticholinergic or antimuscarinic agents.

[0131] The dosages of the compounds of the invention depend upon a variety
of factors including the particular disease to be treated, the severity
of the symptoms, the route of administration, the frequency of the dosage
interval, the particular compound utilized, the efficacy, toxicology
profile, and pharmacokinetic profile of the compound.

[0132] Advantageously, the compounds of formula (I) can be administered
for example, at a dosage of 0.001 to 1000 mg/day, preferably 0.1 to 500
mg/day.

[0133] When the compounds of formula (I) are administered by inhalation
route, they are preferably given at a dosage of 0.001 to 500 mg/day,
preferably 0.1 to 200 mg/day.

[0135] Other features of the invention will become apparent in the course
of the following descriptions of exemplary embodiments which are given
for illustration of the invention and are not intended to be limiting
thereof.

EXAMPLES

[0136] Unless otherwise noted, all starting materials were obtained from
commercial suppliers and used without any further purification; all
reactions are conducted under an inert atmosphere and in dry solvents.

[0138] In a first flask, (R)-quinuclidin-3-amine dihydrochloride (500 mg,
2.51 mmol) was dissolved in MeOH (25 ml) and water (2.50 ml). Sodium
bicarbonate (211 mg, 2.51 mmol) was added, and the reaction was stirred
at room temperature for 1 hour. The reaction was then evaporated to
dryness. The residue was dissolved in dry DMF (25.0 ml), and CDI (407 mg,
2.51 mmol) was added. The reaction was stirred at room temperature for 16
hours.

[0139] In a second flask, bis(3-fluorophenyl)methanol (1.11 g, 5.02 mmol)
was dissolved in dry DMF (25 ml) and treated portionwise with sodium
hydride (60% dispersion in mineral oil, 201 mg, 5.02 mmol) at 0°
C. This second reaction was stirred at room temperature for 30 minutes
and then poured into the first flask.

[0140] The resulting reaction was stirred at room temperature for 2 days.
Then the reaction was portioned between Et2O and water. The organic
phase was dried over Na2SO4, filtered and evaporated to
dryness. The crude was purified by flash chromatography (DCM/MeOH=9/1 to
DCM/MeOH=75/25+0.5% TEA) to obtain (R)-bis(3-fluorophenyl)methyl
quinuclidin-3-ylcarbamate (419 mg).

[0143] In a first flask, (R)-quinuclidin-3-amine dihydrochloride (0.10 g,
0.50 mmol) was dissolved in MeOH (5 ml) and water (0.5 ml). Sodium
bicarbonate (84.0 mg, 1.00 mmol) was added, and the reaction was stirred
at room temperature for 1 hour. The reaction was then evaporated to
dryness. The solid was dissolved in dry DMF (5 ml), and CDI (81.0 mg,
0.50 mmol) was added. The reaction was stirred at room temperature for 7
hours.

[0144] In a second flask, diphenylmethanol (0.18 g, 1.00 mmol) was
dissolved in dry DMF (5 ml) and treated portionwise with NaH (40.0 mg,
1.00 mmol) at 0° C. The reaction was stirred at room temperature
for 20 minutes and then poured into the first flask.

[0145] The resulting reaction was stirred at room temperature overnight.
The reaction mixture was portioned between Et2O and water. The
organic phase was washed with brine, dried over Na2SO4,
filtered, and evaporated to dryness. The crude was purified by flash
chromatography (DCM/MeOH=9/1 to DCM/MeOH=75/25+0.5% TEA) to obtain
(R)-benzhydryl quinuclidin-3-ylcarbamate (105 mg).

[0148] In a first flask, (R)-quinuclidin-3-amine dihydrochloride (50.0 mg,
0.25 mmol) was dissolved in MeOH (2.5 ml) and water (0.25 ml). Sodium
bicarbonate (42.0 mg, 0.50 mmol) was added, and the reaction was stirred
at room temperature for 1 hour. The reaction was then evaporated to
dryness. The solid was dissolved in dry DMF (2.50 ml), and CDI (40.5 mg,
0.25 mmol) was added. The reaction was stirred at room temperature for 7
hours.

[0149] In a second flask, bis(4-fluorophenyl)methanol (111 mg, 0.50 mmol)
was dissolved in dry DMF (5 ml), and sodium hydride (20.0 mg, 0.50 mmol)
was added portionwise at 0° C. The reaction was stirred at
0° C. for 5 minutes and then poured into the first flask, cooled
at 0° C.

[0150] The resulting reaction mixture was stirred at room temperature
overnight. The reaction mixture was then portioned between Et2O and
water. The organic phase was dried over Na2SO4, filtered, and
evaporated to dryness. The crude was purified by flash chromatography
(DCM/MeOH=9/1 to DCM/MeOH=75/25+0.5% TEA) to obtain
(R)-bis(4-fluorophenyl)methyl quinuclidin-3-ylcarbamate (37.0 mg).

[0152] The following compounds were prepared following the route described
in Example 3, using the suitable alcohols instead of
bis(4-fluorophenyl)methanol. These compounds were achieved as a mixture
of diastereoisomers.

[0168] 5-(2-Bromoacetyl)thiophene-2-carbonitrile (33.4 mg, 0.14 mmol) was
added to a solution of (R)-bis(3-fluorophenyl)methyl
quinuclidin-3-ylcarbamate (54 mg, 0.14 mmol, prepared as in example 1) in
ethyl acetate (2 ml). The resulting solution was stirred at room
temperature for two days, then 5-(2-bromoacetyl)thiophene-2-carbonitrile
(3.4 mg, 0.015 mmol) was added, and the stirring was kept for additional
16 hours. Et2O was added and the precipitate was recovered by
filtration to afford
(R)-3-((bis(3-fluorophenyl)methoxy)carbonylamino)-1-(2-(5-cyanothiophen-2-
-yl)-2-oxoethyl)-1-azoniabicyclo[2.2.2]octane bromide (68 mg).

[0172] 2-Bromo-1-(pyridin-2-yl)ethanone hydrobromide (39.2 mg, 0.14 mmol)
was added to a solution of (R)-bis(3-fluorophenyl)methyl
quinuclidin-3-ylcarbamate (52 mg, 0.14 mmol, prepared as in example 1) in
ethyl acetate (2 ml). The reaction was stirred at room temperature for 16
hours, then Et2O (1 mL) was added, and the precipitate was collected
by suction filtration and dried under vacuum at 40° C. overnight.
The product was further purified by preparative HPLC to obtain
(R)-3-((bis(3-fluorophenyl)methoxy)-carbonylamino)-1-(2-oxo-2-(pyridin-2--
yl)ethyl)-1-azoniabicyclo[2.2.2]octane bromide (16.5 mg).

[0181] 2-Chloro-1-phenylethanone (20.8 mg, 0.13 mmol) was added to a
solution of (R)-bis(3-fluorophenyl)methyl quinuclidin-3-ylcarbamate (50
mg, 0.13 mmol, prepared as in example 1) in ethyl acetate (2 ml). The
reaction was stirred at room temperature overnight, then the solvent was
evaporated under vacuum. The residue was treated with Et2O (4 ml)
and sonicated to obtain a solid, which was collected by suction
filtration and purified by flash chromatography (DCM/MeOH=95/5), to
obtain (R)-3-((bis(3-fluorophenyl)methoxy)carbonylamino)-1-(2-oxo-2-pheny-
lethyl)-1-azoniabicyclo[2.2.2]octane chloride (37.2 mg).

[0185] (R)-Bis(3-fluorophenyl)methyl quinuclidin-3-ylcarbamate (50 mg,
0.13 mmol, prepared as in example 1) was added to a solution of
(2-bromoethoxy)benzene (27.0 mg, 0.13 mmol) in ethyl acetate (2 ml). The
mixture was stirred at room temperature overnight. Then
(2-bromoethoxy)benzene (27 mg, 0.13 mmol) was added, and the mixture was
heated at 100° C. for 15 minutes under microwave irradiation. A
catalytic amount of potassium iodide was added, and the reaction was
heated at 120° C. for 1 hour under microwave irradiation. The
precipitate was collected by filtration and dried to obtain
(R)-3-((bis(3-fluorophenyl)methoxy)carbonylamino)-1-(2-phenoxyethyl)-1-az-
oniabicyclo[2.2.2]octane bromide (17 mg).

[0189] To a solution of (R)-bis(3-fluorophenyl)methyl
quinuclidin-3-ylcarbamate (53 mg, 0.14 mmol, prepared as in example 1) in
ethyl acetate (2 ml), 5-(2-bromoethyl)-2,3-dihydrobenzofuran (32.3 mg,
0.14 mmol) was added. The resulting mixture was stirred at room
temperature for 8 days, and then the solvent was evaporated, and the
crude was purified by flash chromatography (DCM/MeOH=95/5) to collect
(R)-3-((bis(3-fluorophenyl)-methoxy)carbonylamino)-1-(2-(2,3-dihydrobenzo-
furan-5-yl)ethyl)-1-azoniabicyclo[2.2.2]octane bromide (22.2 mg).

[0193] (R)-bis(3-Fluorophenyl)methyl quinuclidin-3-ylcarbamate (55 mg,
0.14 mmol, prepared as in example 1) was dissolved in ethyl acetate (2
ml), and 1-(2-bromoethyl)-4-fluorobenzene (21 μl, 0.15 mmol) was
added. The reaction was stirred at room temperature for 24 hours, then
1-(2-bromoethyl)-4-fluorobenzene (6.21 μl, 0.04 mmol) was added again.
After being stirred at room temperature for 2 days, the reaction was
concentrated under vacuum, and the crude was first purified by flash
chromatography (DCM/MeOH=9/1) and then triturated with DCM/Et2O
(1/1) to obtain
(R)-3-((bis(3-fluorophenyl)-methoxy)carbonylamino)-1-(4-fluorophenethyl)--
1-azoniabicyclo[2.2.2]octane bromide (45 mg).

[0197] 2-Chloro-1-phenylethanone (55.0 mg, 0.36 mmol) was added to a
solution of (R)-benzhydryl quinuclidin-3-ylcarbamate (100 mg, 0.30 mmol,
prepared as in example 2) in ethyl acetate (4 ml). The reaction was
stirred at room temperature overnight. The suspension was decanted,
solvent was removed, and the product was evaporated to dryness. Et2O
was added, and the suspension was sonicated. The precipitate was
collected by suction filtration and dried under vacuum at 40° C.
to obtain (R)-3-(benzhydryloxycarbonylamino)-1-(2-oxo-2-phenylethyl)-1-az-
oniabicyclo-[2.2.2]octane chloride (110 mg)

[0201] 2-Chloro-1-phenylethanone (13 mg, 0.09 mmol) was added to a
solution of (R)-bis(4-fluorophenyl)methyl quinuclidin-3-ylcarbamate (32.0
mg, 0.09 mmol, prepared as in example 3) in ethyl acetate (2 ml). The
reaction was stirred at room temperature overnight. Then more
2-chloro-1-phenylethanone (2.7 mg, 0.02 mmol) was added to the solution,
and the reaction was stirred for additional 24 hours. The reaction was
evaporated to dryness, and the residue was triturated in Et2O. Then
isopropyl ether was added, and the product was sonicated and the
precipitate was collected by suction filtration to obtain
(R)-3-((bis(4-fluorophenyl)methoxy)carbonylamino)-1-(2-oxo-2-phenylethyl)-
-1-azoniabicyclo[2.2.2]octane chloride (32.4 mg)

[0205] 2-Chloro-1-(thiophen-2-yl)ethanone (12.0 mg, 0.07 mmol) was added
to a solution of (4-methoxyphenyl)(phenyl)methyl
(R)-quinuclidin-3-ylcarbamate (22.0 mg, 0.06 mmol, prepared as in example
3) in ethyl acetate (2 ml). The reaction was stirred at room temperature
for two days. The reaction was evaporated to dryness. The crude was
purified by flash chromatography (DCM/MeOH=95/5) to obtain
(3R)-3-(((4-methoxyphenyl)(phenyl)methoxy)carbonylamino)-1-(2-oxo-2-(thio-
phen-2-yl)ethyl)-1-azoniabicyclo[2.2.2]octane chloride (23 mg, mixture of
diastereoisomers).

[0208] The following compounds were prepared following the route described
in Example 17 using the suitable intermediates (Example 3, Table 1)
instead of (4-methoxyphenyl)(phenyl)methyl (R)-quinuclidin-3-ylcarbamate.
These compounds were obtained as a mixture of diastereoisomers.

[0210] In a first flask, (R)-quinuclidin-3-amine dihydrochloride (100 mg,
0.50 mmol) was dissolved in MeOH (5 ml), and water (0.5 ml). NaHCO3
(84.0 g, 1.00 mmol) was added, and the reaction was stirred at room
temperature for 1 hour. The reaction mixture was evaporated, and the
residue was dissolved in dry DMF (5 ml), and CDI (81.0 mg, 0.50 mmol) was
added. The reaction was stirred at room temperature overnight.

[0211] In a second flask, phenyl(4-(trifluoromethyl)phenyl)methanol (0.25
g, 1.00 mmol) was dissolved in dry DMF (3 ml), and sodium hydride (60%
dispersion in mineral oil, 24.0 mg, 1.00 mmol) was added portionwise at
0° C. The reaction was stirred at room temperature for 15 minutes
and then was poured into the first flask.

[0212] The resulting mixture was stirred at room temperature overnight,
then it was portioned between Et2O and water. The organic phase was
washed with brine and dried over sodium sulphate. The solvent was removed
in vacuo and the crude was purified by flash chromatography
(DCM/MeOH=95/5 to DCM/MeOH/TEA=75/25/0.5) to obtain
phenyl(4-(trifluoromethyl)-phenyl)methyl (R)-quinuclidin-3-ylcarbamate
(56.0 mg, mixture of diastereoisomers).

[0217] In a first flask, (R)-quinuclidin-3-amine dihydrochloride (80.0 mg,
0.40 mmol) was dissolved in MeOH (8 ml) and water (0.8 ml). Sodium
bicarbonate (67.5 mg, 0.80 mmol) was added, and the reaction was stirred
at room temperature for 1 hour. The reaction was evaporated to dryness,
and the solid was dissolved in dry DMF (8 ml). CDI (65.1 mg, 0.40 mmol)
was added, and the reaction was stirred at room temperature overnight.

[0218] In a second flask, (2-chlorophenyl)(4-chlorophenyl)methanol (100
mg, 0.39 mmol) was dissolved in dry DMF (3 ml) and treated with sodium
hydride (60% dispersion in mineral oil, 15.8 mg, 0.39 mmol) at 0°
C. The ice-bath was removed, and the reaction was stirred at room
temperature for 20 minutes, and then it was added to the first reaction.

[0219] The resulting mixture was stirred at room temperature overnight,
and then it was portioned between Et2O and water. The organic phase
was washed with brine, dried over Na2SO4, filtered, and
evaporated to dryness. The crude was purified by flash chromatography
(DCM/MeOH=9/1 to DCM/MeOH=75/25) to obtain
(2-chlorophenyl)(4-chlorophenyl)methyl (R)-quinuclidin-3-ylcarbamate (95
mg, mixture of diastereoisomers).

[0220] To a solution of (2-chlorophenyl)(4-chlorophenyl)methyl
(R)-quinuclidin-3-ylcarbamate (95.0 mg, 0.23 mmol) in ethyl acetate (3
ml), 2-chloro-1-(thiophen-2-yl)ethanone (37.6 mg, 0.23 mmol) was added,
and the mixture was stirred at room temperature overnight. The solvent
was removed under vacuum, and the residue was taken up in diethyl ether
and filtered. The product was purified by flash chromatography
(DCM/MeOH=95/5) to obtain
(3R)-3-(((2-chlorophenyl)(4-chlorophenyl)methoxy)carbonylamino)-1-(2-oxo--
2-(thiophen-2-yl)ethyl)-1-azoniabicyclo[2.2.2]octane chloride (45.0 mg,
mixture of diastereoisomers).

[0224] In a first flask, (R)-quinuclidin-3-amine dihydrochloride (100 mg,
0.50 mmol) was dissolved in MeOH (10 ml) and water (1 ml). Sodium
bicarbonate (84 mg, 1.00 mmol) was added, and the reaction was stirred at
room temperature for 1 hour. The reaction was evaporated to dryness, and
the solid was dissolved in dry DMF (10 ml). CDI (81 mg, 0.50 mmol) was
added, and the reaction was stirred at room temperature overnight.

[0225] In a second flask, thiophen-2-ylmethanol (114 mg, 1.00 mmol) was
dissolved in dry DMF (3 ml) and treated with sodium hydride (60%
dispersion in mineral oil, 40.0 mg, 1.00 mmol) at 0° C. The
ice-bath was removed, and the reaction was stirred at room temperature
for 20 minutes, and then it was poured into the first flask.

[0226] The resulting mixture was stirred at room temperature overnight,
and then it was portioned between Et2O and water. The organic phase
was washed with brine, dried over Na2SO4, filtered, and
evaporated to dryness. The crude was purified by flash chromatography
(DCM/MeOH=9/1 to 75/25) to obtain (R)-thiophen-2-ylmethyl
quinuclidin-3-ylcarbamate (50 mg).

[0231] In a first flask, (R)-quinuclidin-3-amine dihydrochloride (500 mg,
2.51 mmol) was dissolved in MeOH (25 ml) and water (2.5 ml). Sodium
bicarbonate (211 mg, 2.51 mmol) was added, and the reaction was stirred
at room temperature for 1 hour. The solvent was evaporated to dryness,
and the residue was suspended in dry DMF (25 ml) and treated with
di(1H-imidazol-1-yl)methanethione (448 mg, 2.51 mmol). The reaction was
stirred 4 hours at room temperature.

[0232] In a second flask, NaH (60% dispersion in mineral oil, 187 mg, 4.68
mmol) was added portionwise to a solution of bis(3-fluorophenyl)methanol
(1.03 g, 4.68 mmol) in dry DMF (25 ml) at 0° C. The reaction was
stirred at room temperature for 30 minutes, and then it was added to the
first flask.

[0233] The resulting mixture was stirred at room temperature overnight,
and then the reaction was portioned between Et2O and water. The
organic phase was dried over Na2SO4, filtered, and evaporated
to dryness. The crude was purified by flash chromatography (DCM/MeOH=9/1
and then DCM/MeOH=75/25+0.5% TEA) to obtain
(R)--O-bis(3-fluorophenyl)methyl quinuclidin-3-ylcarbamothioate (60 mg).

[0238] In a first flask, CDI (184 mg, 1.135 mmol) was added to a solution
of (R)-1-benzylpyrrolidin-3-amine (200 mg, 1.13 mmol) in dry DMF (20 ml),
and the reaction was stirred at room temperature overnight.

[0239] In a second flask, bis(3-fluorophenyl)methanol (375 mg, 1.70 mmol)
was dissolved in dry DMF (9 ml) and treated with sodium hydride (60%
dispersion in mineral oil, 68.1 mg, 1.703 mmol) at 0° C. The
ice-bath was removed, and the reaction was stirred at room temperature
for 20 minutes, and then it was added to the first flask.

[0240] The resulting mixture was stirred at room temperature overnight,
and then portioned between Et2O and water. The organic phase was
washed with brine, dried over Na2SO4, filtered, and evaporated
to dryness. The crude was purified by flash chromatography
(DCM/MeOH=98/2) to give (R)-bis(3-fluorophenyl)methyl
1-benzylpyrrolidin-3-ylcarbamate (300 mg).

[0241] To a solution of (R)-bis(3-fluorophenyl)methyl
1-benzylpyrrolidin-3-ylcarbamate (300 mg, 0.71 mmol) in EtOAc (9 ml),
iodomethane (44.2 μl, 0.71 mmol) was added, and the reaction was
stirred at room temperature overnight. The solvent was then removed under
vacuum, and the residue was triturated with diethyl ether. The product
was purified by flash chromatography (DCM/MeOH=99/1) and then by
preparative HPLC to give
(3R)-1-benzyl-3-((bis(3-fluorophenyl)methoxy)carbonylamino)-1-methylpyrro-
lidinium iodide (25 mg).

[0248] The following compounds were prepared following the route described
in Example 23 using the suitable alkylating agents instead of
2-bromo-1-phenylethanone. These compounds were obtained as a racemic
mixture.

[0253] The following compounds were prepared following the route described
in Example 24 using the suitable alkylating agents instead of
2-bromo-1-p-tolylethanone. These compounds were obtained as a racemic
mixture.

[0270] The following compounds were prepared following the route described
in Example 28 using the suitable alkylating agents instead of
2-bromo-1-(2-fluorophenyl)ethanone. These compounds were obtained as a
racemic mixture.

[0294] To a solution of benzhydryl quinuclidine-3-carboxylate (54 mg, 0.17
mmol) in acetonitrile (5 ml), 2-bromo-1-(thiophen-2-yl)ethanone (37.9 mg,
0.18 mmol) was added, and the reaction was stirred at room temperature
for 15 hours. The solvent was evaporated, and the resulting crude was
first purified by preparative HPLC and then by flash chromatography
(DCM/MeOH=95/5 to 9/1) to obtain
3-(benzhydryloxycarbonyl)-1-(2-oxo-2-(thiophen-2-yl)ethyl)-1-azoniabicycl-
o[2.2.2]octane 2,2,2-trifluoroacetate (53.8 mg, racemic mixture).

[0298] Quinuclidine-3-carboxylic acid hydrochloride (150 mg, 0.78 mmol),
EDC (225 mg, 1.17 mmol), and HOBT (180 mg, 1.17 mmol) were dissolved in
dry THF (7.5 ml). (4-Methoxyphenyl)(phenyl)methanol (184 mg, 0.86 mmol)
and TEA (382 μl, 2.74 mmol) were added, and the resulting reaction was
stirred at room temperature for four days. THF was removed under vacuum,
and the crude residue was partitioned between EtOAc and water. The
organic phase was washed with a sat. NaHCO3, dried over
Na2SO4, filtered, and evaporated. The crude was purified by
flash chromatography (EtOAc/MeOH=8/2 to EtOAc/MeOH=7:3+1% NH4OH) to
give (4-methoxyphenyl)(phenyl)methyl quinuclidine-3-carboxylate (37 mg,
mixture of diastereoisomers). The compound was used as such in the next
step.

[0303] Quinuclidine-3-carboxylic acid hydrochloride (0.1 g, 0.52 mmol),
EDC (150 mg, 0.78 mmol), and HOBT (120 mg, 0.78 mmol) were dissolved in
dry THF (3 ml). (2-Chlorophenyl)(4-chlorophenyl)methanol (145 mg, 0.57
mmol) was dissolved in dry THF (2 ml) and then added to the reaction
mixture. Triethylamine (253 μl, 1.83 mmol) was added, and the
resulting reaction was stirred at room temperature overnight. The mixture
was evaporated, and the crude was taken up with EtOAc and washed with
water and brine. The organic phase was separated, dried (sodium sulfate),
filtered, and evaporated under vacuum. The crude was purified by flash
chromatography (DCM/MeOH=95/5) to afford
(2-chlorophenyl)(4-chlorophenyl)methyl quinuclidine-3-carboxylate (40 mg,
mixture of diastereoisomers). The compound was used as such in the next
step.

[0308] Quinuclidine-3-carboxylic acid hydrochloride (80 mg, 0.42 mmol),
EDC (120 mg, 0.62 mmol), and HOBT (96 mg, 0.62 mmol) were dissolved in
dry THF (3 ml). (3,4-Difluorophenyl)(phenyl)methanol (101 mg, 0.46 mmol)
was dissolved in dry THF (2 ml) and then added to the reaction mixture.
Triethylamine (203 μl, 1.46 mmol) was finally added, and the resulting
reaction was stirred at room temperature overnight. The volatiles were
evaporated, and the crude was taken up with EtOAc and washed with water
and then with sat. NaHCO3. The organic phase was dried over sodium
sulfate, filtered, and evaporated under vacuum. The crude was purified by
flash chromatography (DCM/MeOH=9/1) to obtain
(3,4-difluorophenyl)(phenyl)-methyl quinuclidine-3-carboxylate (65 mg,
mixture of diastereoisomers).

[0309] To a solution of (3,4-difluorophenyl)(phenyl)methyl
quinuclidine-3-carboxylate (65 mg, 0.18 mmol) in ethyl acetate (3 ml),
2-chloro-1-(thiophen-2-yl)ethanone (29.2 mg, 0.18 mmol) was added, and
the mixture was stirred at room temperature overnight. The solvent was
removed under vacuum, and the residue was taken up with diethyl ether and
filtered to give
3-(((3,4-difluorophenyl)(phenyl)methoxy)carbonyl)-1-(2-oxo-2-(thiophen-2--
yl)ethyl)-1-azoniabicyclo[2.2.2]octane chloride (20 mg, mixture of
diastereoisomers).

[0313] Quinuclidine-3-carboxylic acid hydrochloride (95 mg, 0.50 mmol),
EDC (143 mg, 0.74 mmol), HOBT (114 mg, 0.74 mmol), and triethylamine (242
μl, 1.73 mmol) were dissolved in dry THF (5 ml).
(3-Fluorophenyl)methanol (68.8 mg, 0.54 mmol) was dissolved in dry THF (2
ml) and added to the reaction mixture. The resulting reaction was stirred
at room temperature overnight. The volatiles were evaporated, and the
crude was taken up with EtOAc and washed with water and then with sat.
NaHCO3. The organic phase was separated, dried over sodium sulfate,
filtered, and evaporated under vacuum to obtain 3-fluorobenzyl
quinuclidine-3-carboxylate (120 mg, racemic mixture). The compound was
used in the next step without any further purification.

[0314] 2-Chloro-1-(thiophen-2-yl)ethanone (67 mg, 0.42 mmol) was added to
a solution of 3-fluorobenzyl quinuclidine-3-carboxylate (100 mg, 0.38
mmol) in a mixture of EtOAc (1.2 ml) with some drops of DMF. The reaction
was stirred at room temperature overnight. Then a second portion of
2-chloro-1-(thiophen-2-yl)ethanone (67 mg, 0.42 mmol) was added, and the
reaction stirred at room temperature for 3 hours. The solvent was
removed, and the residue was dissolved in acetonitrile and heated under
microwave irradiation at 100° C. for 2 hours. Acetonitrile was
evaporated and the crude was triturated with Et2O and then purified
by preparative HPLC to obtain
3-((3-fluorobenzyloxy)carbonyl)-1-(2-oxo-2-(thiophen-2-yl)ethyl)-1-azonia-
bicyclo-[2.2.2]octane 2,2,2-trifluoroacetate (70 mg, racemic mixture).

[0319] 2-Chloro-1-(thiophen-2-yl)ethanone (55 mg, 0.344 mmol) was added to
a solution of 9H-fluoren-9-yl quinuclidine-3-carboxylate (100 mg, 0.31
mmol) in acetonitrile (1 ml). The reaction was stirred at room
temperature overnight. 2-Chloro-1-(thiophen-2-yl)ethanone (55 mg, 0.34
mmol) was added again, and the mixture stirred at room temperature for 3
hours. The solvent was removed, and the residue was re-dissolved in
acetonitrile and heated under microwave irradiation at 100° C. for
3 hours. Acetonitrile was removed, and the residue was triturated with
Et2O and then purified by preparative HPLC to obtain
3-((9H-fluoren-9-yloxy)carbonyl)-1-(2-oxo-2-(thiophen-2-yl)ethyl)-1-azoni-
abicyclo[2.2.2]octane 2,2,2-trifluoroacetate (46 mg, racemic mixture).

[0323] 1-Methylpiperidine-4-carboxylic acid hydrochloride (250 mg, 1.39
mmol), EDC (400 mg, 2.09 mmol), and HOBT (320 mg, 2.09 mmol) were
dissolved in dry THF (10 ml). Bis(3-fluorophenyl)methanol (337 mg, 1.53
mmol) was dissolved in dry THF (4 ml) and then added to the reaction
mixture. Triethylamine (6790, 4.87 mmol) was finally added, and the
resulting reaction was stirred at room temperature overnight. The
reaction was evaporated, and the residue was taken up with EtOAc and
washed with water and sat.NaHCO3. The organic phase was dried over
sodium sulfate, filtered, and evaporated under vacuum. The crude was
purified by flash chromatography (DCM/MeOH=9/1) to afford
bis(3-fluorophenyl)methyl 1-methylpiperidine-4-carboxylate (150 mg).

[0324] To a solution of bis(3-fluorophenyl)methyl
1-methylpiperidine-4-carboxylate (150 mg, 0.43 mmol) in ethyl acetate (5
ml), 2-chloro-1-(thiophen-2-yl)ethanone (69.8 mg, 0.43 mmol) was added,
and the mixture was stirred at room temperature overnight. The solvent
was removed under vacuum, and the residue was taken up in diethyl ether,
filtered, and purified by flash chromatography (DCM/MeOH=95/5) to obtain
4-((bis(3-fluorophenyl)methoxy)carbonyl)-1-methyl-1-(2-oxo-2-(thiophen-2--
yl)ethyl)piperidinium chloride (18 mg).

[0332] To a solution of bis(3-fluorophenyl)methyl
1-methylpiperidine-4-carboxylate (106 mg, 0.31 mmol, prepared as in
example 40) in EtOAc (3 ml), 2-bromo-1-(thiazol-2-yl)ethanone (63.2 mg,
0.31 mmol) was added. The mixture was stirred at room temperature for 27
hours. The solvent was evaporated, and the crude was triturated with
Et2O and collected by filtration. The compound was purified by flash
chromatography (DCM/MeOH=95/5 to 9/1) to obtain
4-((bis(3-fluorophenyl)methoxy)carbonyl)-1-methyl-1-(2-oxo-2-(thiazol-2-y-
l)ethyl)piperidinium bromide (120 mg).

[0336] Oxalyl dichloride (141 μl, 1.67 mmol) was added dropwise to a
solution of 1-methylpiperidine-4-carboxylic acid hydrochloride (300 mg,
1.67 mmol) in DCM (20 ml) and few drops of DMF (catalytic amount). The
reaction was stirred at room temperature for 2 hours. The volatiles were
removed under vacuum, and the crude was taken up with pyridine (20 ml).
1,2-Diphenylethanol (331 mg, 1.67 mmol) was added, and the resulting
suspension was heated under microwave irradiation at 140° C. for 1
hour. Pyridine was removed under vacuum, and the crude was purified by
flash chromatography (DCM/MeOH=97/3 to 94/6) to obtain 1,2-diphenylethyl
1-methylpiperidine-4-carboxylate (200 mg).

[0343] Oxalyl dichloride (236 μl, 2.78 mmol) was added dropwise to a
solution of 1-methylpiperidine-4-carboxylic acid hydrochloride (500 mg,
2.78 mmol) in DCM (30 ml) and few drops of DMF (catalytic amount). The
reaction was stirred at room temperature for 2 hours. The volatiles were
removed under vacuum, and the crude was taken up with pyridine (40 ml).
1,2-Diphenylethanone (546 mg, 2.78 mmol) was added, and the resulting
suspension was heated under microwave irradiation at 140° C. for 1
hour. Pyridine was removed under vacuum, and the crude was purified by
flash chromatography (DCM/MeOH=95/5) to obtain (E)-1,2-diphenylvinyl
1-methylpiperidine-4-carboxylate (62 mg).

[0344] 2-Bromo-1-phenylethanone (57.6 mg, 0.29 mmol) was added to a
solution of (E)-1,2-diphenylvinyl 1-methylpiperidine-4-carboxylate (62
mg, 0.19 mmol) in acetonitrile (3 ml). The reaction was stirred at room
temperature overnight, and then the solvent was evaporated. The crude was
purified by flash chromatography (DCM/MeOH=98/2) and then by flash
chromatography (DCM/MeOH=99/1) to obtain
(1R,4R)-4-(((E)-1,2-diphenylvinyloxy)carbonyl)-1-methyl-1-(2-oxo-2-phenyl-
ethyl)piperidinium bromide (18 mg).

[0348] A mixture of quinuclidine-4-carboxylic acid hydrochloride (100 mg,
0.52 mmol) and thionyl chloride (500 μl, 6.85 mmol) was refluxed for 2
hours. The reaction was cooled to room temperature, and the solvent was
accurately removed. The residue was suspended in dry DCM and treated with
(3-fluorophenyl)methanol (65.8 mg, 0.52 mmol). The reaction was stirred
at room temperature for 24 hours. The solvent was evaporated, and the
residue was dissolved in water (1 ml), basified with NaHCO3 and
extracted twice with EtOAc. The combined organic layers were dried over
Na2SO4, filtered, and evaporated to obtain 3-fluorobenzyl
quinuclidine-4-carboxylate (41 mg, 29.8% yield), which was used in the
next step without any further purification.

[0353] Quinuclidine-3-carboxylic acid hydrochloride (55 mg, 0.29 mmol),
EDC (83 mg, 0.43 mmol), and HOBT (65.9 mg, 0.43 mmol) were dissolved in
dry THF (5 ml) under a nitrogen atmosphere. Triethylamine (140 μl,
1.00 mmol) and phenylmethanethiol (37.1 μl, 0.32 mmol) were dissolved
in dry THF (2 ml) and then added to the reaction mixture. The resulting
suspension was stirred at room temperature overnight. The mixture was
evaporated, and the crude partitioned between EtOAc and water. The
organic phase was washed with saturated NaHCO3, dried over sodium
sulfate, filtered, and evaporated under vacuum to obtain S-benzyl
quinuclidine-3-carbothioate (110 mg, crude), which was used in the next
step without any further purification.

[0354] 2-Chloro-1-(thiophen-2-yl)ethanone (68.0 mg, 0.42 mmol) was added
to a solution of S-benzyl quinuclidine-3-carbothioate (100 mg, 0.38 mmol)
in acetonitrile (1 ml). The reaction was stirred at room temperature
overnight, then a second portion of 2-chloro-1-(thiophen-2-yl)ethanone
(68.0 mg, 0.42 mmol) was added, and the reaction was stirred at room
temperature for additional 3 hours. The solvent was removed in vacuo, and
the residue was dissolved in acetonitrile and heated under microwave
irradiation at 100° C. for 3 hours. Acetonitrile was removed, and
the residue was triturated with Et2O. The crude was then purified by
a preparative HPLC (eluent: CH3CN/H2O) to obtain
3-(benzylthiocarbonyl)-1-(2-oxo-2-(thiophen-2-yl)ethyl)-1-azoniabicyclo[2-
.2.2]octane chloride (8.9 mg, 5.5% yield).

[0365] The interaction with M3 muscarinic receptors can be estimated by
the results of in vitro studies which evaluated the M3/M2 binding assays,
the potency of the test compounds and the offset of the inhibitory
activity produced after washout of the antagonists in isolated guinea pig
trachea and by the in vivo duration of action against
acetylcholine-induced bronchospasm in the guinea pig.

[0367] The day of experiment, frozen membranes were resuspended in buffer
C (50 mM Tris-HCl pH 7.4, 2.5 mM MgCl2, 1 mM EDTA). The non
selective muscarinic radioligand [3H]-N-methyl scopolamine (Mol.
Pharmacol., vol. 45, pp. 899-907, which is incorporated herein by
reference in its entirety) was used to label the M2 and M3 binding sites.
Binding experiments were performed in duplicate (ten point concentrations
curves) in 96 well plates at radioligand concentration of 0.1-0.3 nM. The
non specific binding was determined in the presence of cold N-methyl
scopolamine 10 W. Samples (final volume 0.75 ml) were incubated at room
temperature for 60 minutes for M2 and 90 minutes for M3 binding assay.
The reaction was terminated by rapid filtration through GF/B Unifilter
plates and two washes (0.75 ml) with cold buffer C using a Packard
Filtermate Harvester. Radioactivity on the filters was measured by a
microplate scintillation counter TriCarb 2500 (PerkinElmer).

[0368] The values of inhibitory M3 activity tested on compounds are
comprised between 0.265 and 1514 nM.

Example 48

In Vitro Interaction with the M3 Receptors

[0369] The potency of the antagonist activity in isolated guinea pig
trachea was investigated following a method previously described by
Haddad E B et al. in Br. J. Pharmacol., vol. 127, pp. 413-420 (1999),
which is incorporated herein by reference in its entirety, with few
modifications.

[0370] A cumulative concentration-response curve to test antagonists was
constructed on preparations precontracted by carbachol, until a complete
inhibition of smooth muscle tone was achieved. The concentration of
antagonist producing a 50% reversal of carbachol-induced tonic
contraction (IC50) was taken as a measure of its potency in this
bioassay.

[0371] In the experiments aiming at assessing the offset of the inhibitory
effects produced by test compounds, the minimal concentration of the test
compounds known to produce a maximal inhibitory effect was administered
to carbachol-precontracted preparations. As soon as the tonic contraction
was completely reversed, the organ bath solution was renewed and
preparations were thoroughly washed with fresh Krebs solution. Carbachol
(0.3 μM) was administered again (at 30 minute interval between washout
and next administration) during the next 4 hours.

[0372] After the administration of carbachol, the inhibitory effects of
the compounds of the invention, administered at a concentration of 10 nM,
were expressed as percentage of the recovery of the contracting response
to carbachol. The percentage of recovery four hours after the washout was
lower than 50%.

Example 49

In Vivo Studies

[0373] The in vivo tests on acetylcholine-induced bronchospasm in guinea
pig were performed according to H. Konzett H and Rossler F, Arch. Exp.
Path. Pharmacol., vol.

[0374] 195, pp. 71-74 (1940), which is incorporated herein by reference in
its entirety. Aqueous solutions of the test compounds were instilled
intratracheally in anaesthetised mechanically ventilated guinea pigs.
Bronchial response to intravenous acetylcholine challenge was determined
before and after drug administration, and changes in pulmonary resistance
at several time-points were expressed as percent of inhibition of
bronchospasm.

[0375] The bronchodilator activity of the tested compounds persisted
unchanged up to 24 hours after the administration.

Example 50

Lung Stability

[0376] Firstly, fresh rat lungs (previously, washed in saline) are
homogenized in Amm. Formiate Buffer 20 mM. In order to demonstrate that
the compounds are degraded, stability in lung homogenate at 1 and 5 hours
was tested for the compound of the invention. Briefly 10 μl of a stock
solution 250 μM of the compound in acetonitrile were added to 1 ml of
lung homogenate, and samples were incubated at 37° C. Lung
homogenate (50 μL) was taken after 0, 1 and 5 hours of incubation and
added to 200 μl of acetonitrile with addition of verapamil as internal
standard (250 ng/ml). Samples were analysed by HPLC-MS/MS analysis. Lung
stability is calculated as percentage remaining after 1 and 5 hours by
dividing the peak area at 1 or 5 hours by the area of the peak at time 0.

[0377] More than 79% of tested compounds could be still detected after 1
hour of incubation and more than 57% after 5 hours, indicating these
compounds are stable in presence of homogenated lung.

[0378] Where a numerical limit or range is stated herein, the endpoints
are included. Also, all values and subranges within a numerical limit or
range are specifically included as if explicitly written out.

[0379] Obviously, numerous modifications and variations of the present
invention are possible in light of the above teachings. It is therefore
to be understood that, within the scope of the appended claims, the
invention may be practiced otherwise than as specifically described
herein.

[0380] All patents and other references mentioned above are incorporated
in full herein by this reference, the same as if set forth at length.